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大鼠脑皮质星形胶质细胞的体外优化培养及鉴定
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Journal Article
大鼠脑皮质星形胶质细胞的体外优化培养及鉴定
2015
Overview
目的旨在获得高纯度的星形胶质细胞,并对不同培养时期的细胞进行鉴定,为进一步研究奠定基础。方法常规分离新生SD大鼠大脑皮质并制备单细胞悬液,采用差速黏附加摇床震荡法对所获得的细胞进行纯化。倒置相差显微镜及HE染色观察细胞形态,GFAP免疫荧光对获得细胞进行鉴定。结果原代培养的皮质细胞从3d开始迅速增殖,9-12d即可铺满瓶底,此时细胞呈明显的分层生长,星形胶质细胞位于底层,形态多样,阳性率为（67.2±7.1）%;经过1次传代后,GFAP阳性率有所提升（84.0±6.0）%,但不能完全去除小胶质细胞及少突胶质细胞,细胞仍呈分层生长;至第3次传代后,获得大量几乎为单一种类的星形胶质细胞,其胞体较大、突起较短粗,一般2-3个,胞核圆形或椭圆形,常偏于一侧,GFAP染色呈强阳性,阳性率可达（97.6±2.4）%。结论通过差速黏附与摇床震荡相结合的方法,获得了高纯度且生长状态良好的星形胶质细胞。
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